Polynucleotide amplification techniques such as the polymerase chain reaction (PCR) are widely used to amplify polynucleotides for a wide variety of applications, including structural and biochemical studies and diagnotics techniques. Despite its usefulness, a number of undesired reactions increase the complexity of the polymerase products and necessitate careful purification or other approaches to reduce the impact of the undesired reactions. These reactions include the synthesis of oligonucleotides aborted during the initiation of transcription, the use of alternative template initiation sites, polymerase slippage, and the addition of one or more non-templated nucleotides at the 3′ termini of nascent transcripts (e.g., through polymerase based terminal transferase activity).
What are needed are new approaches to reduce or eliminate undersired reactions or the impact of undersired reactions.